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-Structure paper
Title | New approach for membrane protein reconstitution into peptidiscs and basis for their adaptability to different proteins. |
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Journal, issue, pages | Elife, Vol. 9, Year 2020 |
Publish date | Mar 3, 2020 |
Authors | Gabriella Angiulli / Harveer Singh Dhupar / Hiroshi Suzuki / Irvinder Singh Wason / Franck Duong Van Hoa / Thomas Walz / |
PubMed Abstract | Previously we introduced peptidiscs as an alternative to detergents to stabilize membrane proteins in solution (Carlson et al., 2018). Here, we present 'on-gradient' reconstitution, a new gentle ...Previously we introduced peptidiscs as an alternative to detergents to stabilize membrane proteins in solution (Carlson et al., 2018). Here, we present 'on-gradient' reconstitution, a new gentle approach for the reconstitution of labile membrane-protein complexes, and used it to reconstitute reaction center complexes, demonstrating that peptidiscs can adapt to transmembrane domains of very different sizes and shapes. Using the conventional 'on-bead' approach, we reconstituted proteins MsbA and MscS and find that peptidiscs stabilize them in their native conformation and allow for high-resolution structure determination by cryo-electron microscopy. The structures reveal that peptidisc peptides can arrange around transmembrane proteins differently, thus revealing the structural basis for why peptidiscs can stabilize such a large variety of membrane proteins. Together, our results establish the gentle and easy-to-use peptidiscs as a potentially universal alternative to detergents as a means to stabilize membrane proteins in solution for structural and functional studies. |
External links | Elife / PubMed:32125274 / PubMed Central |
Methods | EM (single particle) |
Resolution | 3.3 - 4.4 Å |
Structure data | EMDB-20950, PDB-6uz2: EMDB-20959: Cryo-EM structure of the mechanosensitive channel MscS reconstituted into peptidiscs EMDB-20962, PDB-6uzl: |
Source |
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Keywords | TRANSLOCASE / membrane protein / ABC transporter / membrane mimetic / peptidisc / TRANSPORT PROTEIN / mechanosensitive channels / MscS |